工程化人源性抗-HBs Fab片段抗原结合位点的研究

取自己制备的纯化人源性抗 HBs Fab 片段。用表达的PreS, PreS1, PreS2 蛋白及合成的PreS1 (21 47aa)27 肽检测其抗原结合位点。结果显示该Fab 片段在非还原状态下分子量约为54kD, 在还原状态下分解为大小两个片段, 大片段为Fab的Fd 链, 分子量约为28kD; 小片段为Fab 的κ链, 分子量约为26kD。该Fab 片段与PreS及PreS1 蛋白呈现结合反应, 说明其抗原结合位点位于HBsAg 的PreS1 区

Study on Epitopes Recognized by An Engineered Human Anti HBs Fab Fragment

To investigate the antigenic sites in HBsAg recognized by an engineered human anti HBs Fab fragment prepared previously,E coli expressing HBV PreS,PreS1,and PreS2 peptides were run in SDS PAGE under reducing and non reducing conditions respectively,and then,Western blotting analysis was performed with the purified Fab fragment In addition,a synthetic peptide of HBV PreS1(residue 21 47) was used in immuno binding assay as well Positive results were found in interactions between PreS and Fab,PreS1 and Fab,as well as the synthetic PreS1 peptide and Fab It is suggested that the epitope recognized by our previously prepared anti HBs Fab appeared to be mapped in HBV PreS1